To develop a non-intravenous, needle-less gene transfer platform for systemic deficiency disorders, including hemophilia and alpha (1)- antitrypsin deficiency, Copernicus Therapeutics has evaluated intratracheal delivery of proprietary formulations of compacted DNA (PLASmin complexes). Our gene transfer system compacts single molecules of plasmid DNA into 15-25 nm particles, and this formulation is capable of efficiently transfecting non-dividing cells. In preliminary studies, intratracheal administration of non-targeted PLASmin complexes generated over 1,000-fold higher levels of transgene expression than naked DNA. Significant levels of human factor IX (-300 ng/ml) in lung epithelial lining fluid were measured following a single intratracheal dose of PLASmin complexes targeted with a ligand for the serpin enzyme complex receptor (SECR), a receptor expressed at the lumenal surface of bronchial epithelium. To advance this technology toward human trials, we propose to: i) identify the cell type transfected following intrapulmonary delivery of non-targeted PLASmin complexes; ii) evaluate SECR-targeted PLASmin complexes for intrapulmonary gene delivery; and iii) determine plasma and bronchoalveolar lavage fluid levels of human factor IX and alpha(1)-antitrypsin after intrapulmonary administration of non-targeted and SECR-targeted PLASmin complexes. Our promising results suggest that patients with systemic genetic deficiency states could be treated with aerosolized PLASmin complexes administered daily or several times per week. PROPOSED COMMERCIAL APPLICATIONS: Although gene therapy holds great promise for treating human disease, limitations in safety and efficacy have restricted its commercial development. Our non-toxic formulation of condensed DNA, termed PLASmin complexes, are stable in physiologic fluids, transfect post- mitotic cells, and generate high level gene expression in lung following an intrapulmonary dose. Delivered as an aerosol via a hand-held inhaler, PLASmin complexes may provide an effective and simple therapy for genetic deficiency states, including hemophilia and alpha(1)-antitrypsin deficiency.